Inflammation plays an important role in acute lung injury (ALI). Hesperetin (HES), a natural flavanone and an aglycone of hesperidin, has established potent anti-inflammatory activity. The aim of this study was to evaluate the potential protective effect of HES on lipopolysaccharide (LPS)-induced ALI in mice and to illuminate its possible directly target. Results indicated that HES pretreatment significantly attenuated LPS-induced pulmonary pathological injury, total protein concentration, markedly decreased the number of neutrophils and the levels of inflammatory cytokines, TNF-α and IL-6, in ALI model in vivo and in vitro. Meanwhile, pretreatment with HES dramatically reduced myeloperoxidase (MPO) activity in LPS-induced ALI mice. Additionally, using molecular docking and co-immunoprecipitation assay, HES showed a directly bind with myeloid differentiation 2 (MD2), in which HES could inhibit MAPK activation, regulate IκB degradation, block the interaction MD2 and its co-receptor Toll-like receptor 4 (TLR4). Taken together, HES showed a significantly protective effect against LPS-induced ALI, which might be associated with MD2 protein. These results attested HES worthy of further progress into an adjunctive potential drug for the treatment for ALI.

The main goal of this study was to evaluate the effects of arbutin (AR) on lipopolysaccharide (LPS)-induced lung injury. A lung injury rat model was established by intravenous LPS administration. We found that levels of inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) in both serum and lung tissue were significant increased after LPS challenge. In addition, pathological conditions were examined in rat lungs, and it was demonstrated that AR-pretreatment reduced LPS-induced malondialdehyde (MDA) levels and increased LPS-induced superoxide dismutase (SOD) activity. Moreover, the expression of sirtuin1 (SIRT1), nuclear erythroid factor 2-related factor 2 (Nrf-2), heme oxygenase-1 (HO-1) as well as the phosphorylation of NF-κBp65 and IκBα were increased with LPS-induced lung injury, and were significantly restored by AR treatment. Together, our results indicated that SIRT1 is a potential therapeutic target in LPS-induced lung injury, and that AR may be a novel therapeutic in patients with acute lung injury.

In order to improve the reproductive capacity of adenosine A2A receptor gene knockout mice and strengthen their breed conservation working, the reproductive capacity of adenosine A2A receptor gene knockout mice was analyzed based on their reproductive record. The results showed that while no significant differences of average litter size were observed in the generations of Fback cros , F1 and F2 (P>0.05), the average litter size of Fback cros , F1 and F2 were all significantly higher than that of F3 (P<0.01). It was indicated that the reproductive capacity of adenosine A2A receptor gene knockout mice decreased with the increase of generations, but their reproductive capacity can be improved by back cross.

The genetic diversity and genetic structure of Wenzhou buffaloes distributed in Pingyang,Ruian,Yongjia,Yueqing and Wenzhou in Zhejiang province were analyzed by ISSR-PCR.A total of 154 polymorphic loci of 186 were amplified using 9 primers selected from 22 primers.Percentage of polymorphism loci(PPL) was 82.80%,observed number of alleles was 1.8420,effective number of alleles was 1.5702,Nei's gene diversity was 0.3108,Shannon's information index was 0.4687,all of the genetic parameters in population level were higher than individual level.Genetic differentiation level(Gst) calculated according to Nei's gene diversity was 0.1840,which suggested that 18.40% genetic variation was present in populations.UPMGA cluster analysis in the strains of Wenzhou buffaloes also demonstrated that Wenzhou buffaloes has lower genetic diversity among the populations and the genetic variation mainly exists in the individual ones.

主要论述了实验动物学在生物医学发展中的重要性,分析了在温州医学院《实验动物学》课程教学中存在的问题,就如何通过提高教师素质、调整教学内容、更新教学手段和完善考核方式等教学改革方案来提高《实验动物学》的教学质量进行了探索,并对该校《实验动物学》课程的教学改革提出了新的要求。

实验动物中心以高校基层党组织作为高校党组织的基础,必须充分发挥基层党组织在学校推动发展、服务师生、凝聚人心、促进和谐的作用,充分发挥基层党组织的战斗堡垒作用,把为广大师生服务作为基层党组织的基本价值取向和功能定位。

To explore the expression changes of leptin and NPY associated with body weight control and energy balance regulation after repeated fasting/refeeding rats given high fat diet,a model of repeating fasting/refeeding rats(repeated cycles of 1 d fasting and 1 d refeeding for 6 week fed with common fat diet)was designed,then the model group was switched to high fat diet everyday(high-fat model group),and the control group was divided into high-fat and common-fat control groups.By the end of 12th week,the concentrations of serum lipid and leptin were measured,the expression of NPY mRNA in aorta were detected by RT-PCR.The results showed that the serum leptin level of model group was significantly higher than that of control group(P0.05),while the expression of NPY mRNA in aorta was significantly lower than that of control group(P0.01);The serum TC,TG,LDL-C,and leptin levels of high-fat model group were significantly lower than those of high-fat control group whereas significantly higher than those of common-fat control group(P0.05),while the expression of NPY mRNA in aorta was significantly higher than that of high-fat and common-fat control groups(P0.01),the lesion degrees of aorta were also lighter relatively.It concluded repeated fasting/refeeding feeding pattern may the leptin system of energy balance to control body weight,can reduce the leptin resistance induced by high fat diet,NPY may the process of energy balance regulation.

To investigate the influence of unilateral testicular torsion on the contralateral testicular spermatogenetic function and the protective effects of safflower Injection(SI)in early stage of contralateral testis injury,21 prepubertal sprague-dawley rats were equally randomized into group A(sham-operation control),B(torsion/detorsion)and C(torsion/detorsion plus injection of SI),with 7 rats in each.The testicular torsion/detorsion model was established by the turner method.All the rats were fed under the same condition for 7 days and sacrificed,and all the right(untwisted)testis were removed from rats.Then the biochemical parameters such as malondialdehyde(MDA),super oxide dlsmutase(SOD)and interleukin 6(IL-6)were evaluated;histology and seminiferous tubule diameters were assessed.Results compared with group B,the level of MDA and IL-6 in group C decreased significantly,while the activity of SOD increased significantly.It was also known that the mean seminiferous tubular diameter of contralateral testis and histological grades maintained in a better condition.Conclusion safflower Injection could reduce the damages on contralateral testis of prepubertal rats with unilateral testicular torsion during the early period,which has relation with anti-free radicaland inhibiting lipid peroxidatic reaction.

建立用于ALK3基因敲除小鼠品种保存的体外受精—胚胎移植(IVF-ET)模型。采用杂合子雌性ALK3小鼠卵子与杂合子雄性ALK3小鼠精子进行体外受精,收集2-cell,进行胚胎冷冻,1周后进行胚胎复苏移植,通过PCR方法进行子代鉴定。结果表明,冻存胚胎320枚,复苏获得存活胚胎96枚,移植80枚,产子23只,获得杂合子11只,通过体外受精—胚胎移植可对ALK3基因敲除小鼠进行保种鉴定。

To obtain the information of characteristics of reproduction and growth of A2a mice and establish the normal hematological indexes and biochemical indexes,the normal Organ weight in SPF A2a knockout mice and C57BL/6 mice.The growth and reproduction performances including litter size,weanling rate,body length,body weight and organ weight were investigated in the A2a mice;20 male and 20 female SPF A2a mice and 20 C57BL/6 mice of 60 days sold were selected.The indexes of hematology,biochemistry and Organ coefficient of SPF A2a mice and that of C57BL/6 mice were measured by automatibiochemical and analyze.Results RDW and PLT of C57BL/6 mic were different from A2a mices(P<0.05),while the ALT and CK were lower in male mice than in females of A2a mice(P<0.05),CK were significant difference(P<0.01).It were differences of organ coefficient.

Objective To analyse genetic polymorphisms and genetic stability in mtDNA D-loop sequence on the fifth generation of A2a mice.Methods Fifty heterozygote mice were selected and reproduced to five generations.mtDNA D-loop region of them were amplified respectively by PCR with two pairs of primers and then sequenced.The result of DNA sequence compared with standard sequence of mice.Result The Sequences of D-loop gene of the Fifty A2a mice ware the same totally,and were the same as the standard sequences of mice.Conclusion The A2a knockout mice colony showed good genetic stability and provide the invaluable resource for experimental animal research as well.

Thirty six SD male rats weighted 80~100 g were randolnly divided into three grougs,and fed on diets as follows:control(Cu 13.15 mg/kg),copper toxic I(Cu 650 mg/kg)and copper toxic Ⅱ(Cu 850 mg/kg)for studying effects of copper toxicity on liver in rats by the methods of experimental pathology for nine weeks for exploring the effect of high diet copper on some liver parameters.The result showed that lesions were appeared in high copper groupsⅠ and Ⅱ when compared those with control group.There were increased the liver of rats coefficien in high copper groupsⅠ and Ⅱ(P0.05).Also,LDH,GOT and liver copper contents in high copper groupsⅠand Ⅱ were much higher in the control group(P0.01).The control group SOD activities were higher in high copper groupⅠand Ⅱ the dietary copper in range of 650~850 mg/kg caused pathological injures in liver of SD rats.

The introduced A2a gene knockout heterozygous mice were fed in SPF environment for propagation according to genetics principles. The genomic DNA from tail tissue of each genotype mice was extracted to identify their genotypes with PCR. The results showed that there were 3 genotypes (wild type, heterozygous and homozygote) in offspring of A2a gene knockout heterozygous mice.

Objective To breed and identify PAX-8 gene knock-out mice. Methods Heterozygote mice were bred and reproduced.Wild type, heterozygote genotype,and homozygote genotype would appear in the offspring of parents. Genome DNA extracted from the mice's tails was subjected to PCR test for genotype identification. Male heterozygote mice were selected to mate with the female heterozygote mice to acquire homozygote baby mice according to Mendel law. Results The breeding and reproducing were successful and more heterozygote genotype mice were reproduced. Conclusions Appropriate methods of breeding, reproducing, and identifying are effective ways to gain PAX-8 gene knock-out mice from heterozygote mice.

为了繁育和鉴定ALK3基因敲除小鼠,将引进的杂合子小鼠进行饲养并繁殖,繁殖成功后提取每种基因型小鼠组织基因组DNA,用PCR法进行鉴定,将其雄性杂合子与雌性杂合子交配.结果表明:ALK3杂合子小鼠的繁殖和饲养均获得成功,亦获得了较多的基因敲除纯合子小鼠.说明正确的饲养繁殖以及鉴定方法是从杂合子中获得ALK3基因敲除纯合子小鼠的有效途径.

We report here the clinical, genetic and molecular characterization of four Chinese families with Leber's hereditary optic neuropathy (LHON). There were variable severity and age-of-onset in visual impairment among these families. Strikingly, there were extremely low penetrances of visual impairment in these Chinese families. Sequence analysis of complete mitochondrial genomes in these pedigrees showed the homoplasmic T3394C (Y30H) mutation, which localized at a highly conserved tyrosine at position 30 of ND1, and distinct sets of mtDNA polymorphisms belonging to haplogroups D4b and M9a. The occurrence of T3394C mutation in these several genetically unrelated subjects affected by visual impairment strongly indicates that this mutation is involved in the pathogenesis of visual impairment. However, there was the absence of functionally significant mtDNA mutations in these four Chinese pedigrees carrying the T3394C mutation. Therefore, nuclear modifier gene(s) or environmental factor(s) may play a role in the phenotypic expression of the LHON-associated T3394C mutation.

目的 探讨二巯基丁二酸(DMSA)与维生素C、蛋氨酸联合应用对染铅ICR小鼠血铅及肝脏的影响. 方法 实验组小鼠饮用0.1%的醋酸铅溶液6周后, 随机分为模型对照组、低剂量DMSA组、高剂量DMSA组及低剂量DMSA与维生素C、蛋氨酸联合应用组(联合组).给药4周后,测小鼠的血铅.并取肝脏组织进行苏木素-伊红(HE)染色,观察肝脏的形态学改变. 结果 低剂量DMSA组、高剂量DMSA组与联合组小鼠血铅浓度和模型对照组差异有显著性( P<0.01); 正常对照组血铅浓度和模型对照组差异有显著性(P <0.01).肝病理学表现为肝细胞可见变性水肿,其中以联合组水肿程度最轻.结论 DMSA与维生素C、蛋氨酸联合应用对小鼠体内铅的排泄具有更好的效果.

目的 测定ALK3/α-MHC和C57BL/6小鼠的生理生化正常值及其脏器重量等.方法 采用全自动生化分析仪及血细胞分析仪分别8～9周龄C57BL/6、ALK3雌、雄小鼠的常用血液学指标及生化指标值进行检测;并测定了各小鼠的脏器.结果 C57BL/6和ALK3/α-MHC小鼠血液的PIT、HGB差异显著(P<0.05).C57BL/6和ALK3/α.MHC雌、雄之间小鼠丙氨酸转氨酶(ALT)、肌酸激酶(CK)差异显著;乳酸脱氢酶(LDH)差异极显著.脏器系数无显著差异.结论 ALK3/α-MHC血常规、血清生化指标和脏器指标的测定数据,将为从事生物医药研究等的科研人员提供参考.

We report here the clinical, genetic, and molecular characterization of three Chinese families with Leber's hereditary optic neuropathy (LHON). There were variable severity and age of onset in visual impairment among these families. Strikingly, there were extremely low penetrances of visual impairment in these Chinese families. Sequence analysis of complete mitochondrial genomes in these pedigrees showed the homoplasmic T14502C (I58V) mutation, which localized at a highly conserved isoleucine at position 58 of ND6, and distinct sets of mtDNA polymorphisms belonging to haplogroups M10a, F1a1, and H2. The occurrence of T14502C mutation in these several genetically unrelated subjects affected by visual impairment strongly indicates that this mutation is involved in the pathogenesis of visual impairment. Here, mtDNA variants I187T in the ND1, A122V in CO1, S99A in the A6, and V254I in CO3 exhibited an evolutionary conservation, indicating a potential modifying role in the development of visual impairment associated with T14502C mutation in those families. Furthermore, nuclear modifier gene(s) or environmental factor(s) may play a role in the phenotypic manifestation of the LHON-associated T14502C mutation in these Chinese families.

目的 分析ICR小鼠线粒体ATPase6和D-Loop基因多态性及封闭群ICR小鼠遗传稳定性.方法 在一个亲本雌鼠100只繁殖产生的共1 000只后代群体中随机取12周龄雄鼠20只,提取小鼠睾丸生殖细胞DNA,PCR扩增小鼠线粒体ATPase6和D-Loop基因,产物纯化后测序,测序结果与小鼠线粒体标准序列比对,比较20只小鼠的序列差异.结果 检测的20只小鼠ATPase6和D-Loop基因序列完全一致,且与小鼠线粒体标准序列完全一致.结论 ICR小鼠群体内未检测到ATPase6和D-Loop基因的多态性变异,该群体具有较好的线粒体遗传稳定性.